Background: Sarcomas are rare, diverse malignancies with limited therapeutic options and poor clinical outcomes. Preclinical models that preserve tumour biology are urgently needed to advance mechanistic understanding and functional precision oncology. We established and comprehensively characterized 29 early-passage patient-derived sarcoma cell (PDC) cultures from 19 patients, representing 11 sarcoma subtypes. Multi-region and multi-site sampling enabled generation of PDCs from spatially distinct areas of individual tumours and from matched primary, recurrent and metastatic lesions.

Methods: PDCs underwent genomic, transcriptomic and proteomic profiling alongside extracellular vesicle (EV) biomarker evaluation and phenotypic and drug-response assays.

Results: Copy-number variant (CNV) analysis revealed recurrent alterations affecting key regulators of cell cycle control and growth signalling. Bulk RNA-sequencing captured substantial inter- and intra-subtype heterogeneity. Proteomic and EV analyses recapitulated subtype- and site-specific differences. A functional drug screen of 38 clinically relevant and investigational agents identified both shared and divergent therapeutic vulnerabilities in the different PDCs.

Discussion: These results demonstrate that early-passage sarcoma PDCs can be regarded as biologically faithful and experimentally tractable models that capture lineage identity, tumour evolution and functional heterogeneity. Integrated multi-omic and functional profiling reveals therapeutic vulnerabilities not evident from genomic data only, supporting PDCs as valuable platforms for translational sarcoma research.